Reverse Genetics for Functional Genomics of Phytopathogenic Fungi and Oomycetes
Figure 2
Schematic representation of a gene replacement strategy. The - and - flanking regions (left and right arms) of the target gene are PCR amplified from genomic DNA of wild-type strain and ligated into vector containing the hygromycin phosphotransferase (hpt) gene. This vector is then linearized using restriction enzyme and introduced into the protoplast of the wild-type strain through transformation. Homologous recombination replaces the gene of interest with hpt gene.
